Difference between revisions of "Part:BBa K3187020"
 
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<partinfo>BBa_K3187020 short</partinfo>
 
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The Sortase A7M is a mutated form of the enzyme Sortase from Staphylococcus aureus. Sortase is an enzyme that catalyzes the formation of a peptide bond between a protein containing a C-terminal LPETG amino acid sequence and a protein containing an N-terminal polyG sequence (two or more glycines on the N- terminus). Peptide bond formation is between the threonine (T) of the LPETG and the N- terminal glycine. Due to the mutations it is calcium-independent which is why it can be used in vivo without disturbing the calcium concentrations in cells. It has a molecular weight of 17,85 kDa.
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                <h3>Profile</h3>
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                <table style=“width:80%“>
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                <tr>
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                <td><b>Name</b></td>
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                <td>Tobacco Vein Mottling Virus (TVMV) Protease Cleavage Site</td>
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                </tr>
  
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                <tr>
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                <td><b>Base pairs</b></td>
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                <td>18</td>
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                </tr>
  
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                <tr>
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                <td><b>Origin</b></td>
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                <td>Tobacco Vein Mottling Virus (TVMV) </td>
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                </tr>
  
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<p style="text-align:center;"><img src="https://static.igem.org/mediawiki/2018/c/c7/T--TU_Darmstadt--NADPHassay.png" alt="ycdW mechanism“ width="150" height="180"></p>
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                <td><b>Properties</b></td>
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<p align="center"><b>Figure 2:</b> Reaction mechanism of YcdW </p> <br>
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                <td> recognition site for the TVMV protease</td>
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===Usage and Biology===
  
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                <p>
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The protease cleavage site from the Tobacco Vein Mottling Virus (TVMV) can be used to specifically cleave a protein at the sequence ETVRFQS.
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<p>
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We used this recognition site to cleave our protein in front of its polyG-tag, so that it has a C-terminal polyG-tag that can be used for sortase-mediated ligation of a GGGG-tagged and a LPETGG-tagged protein.
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<br><br>
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To read more about the sortase-mediated ligation, please have a look at our <a href=https://2019.igem.org/Team:TU_Darmstadt/Project/Sortase>wiki</a>.
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===Usage and Biology===
 
  
 
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===Functional Parameters===
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Latest revision as of 17:11, 21 October 2019


Tobacco Vein Mottling Virus Protease Cleavage Site

Profile

Name Tobacco Vein Mottling Virus (TVMV) Protease Cleavage Site
Base pairs 18
Origin Tobacco Vein Mottling Virus (TVMV)
Properties recognition site for the TVMV protease

Usage and Biology

The protease cleavage site from the Tobacco Vein Mottling Virus (TVMV) can be used to specifically cleave a protein at the sequence ETVRFQS.

We used this recognition site to cleave our protein in front of its polyG-tag, so that it has a C-terminal polyG-tag that can be used for sortase-mediated ligation of a GGGG-tagged and a LPETGG-tagged protein.

To read more about the sortase-mediated ligation, please have a look at our wiki.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]